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利用磁性纳米颗粒和选择反应监测质谱法鉴定果汁/生菜中的细菌。

Identification of bacteria in juice/lettuce using magnetic nanoparticles and selected reaction monitoring mass spectrometry.

机构信息

Department of Chemistry, National Dong Hwa University, Hualien, Taiwan.

出版信息

J Food Drug Anal. 2019 Apr;27(2):575-584. doi: 10.1016/j.jfda.2018.09.006. Epub 2018 Oct 26.

Abstract

Ensuring food safety requires a rapid and reliable method for detecting food-borne pathogens. Mass spectrometry has been demonstrated as a powerful tool to classify pure bacterial species. However, matrix interference from food backgrounds may lead to false results because of the suppression of microbial signals. It is useful to develop a method for bacterial enrichment and marker identification in food samples. Magnetic zirconia nanoparticles were used to concentrate spiked microorganisms from apple juice/lettuce under specific conditions (pH 4.5). Bacterial identification was achieved using nanoLC-MS. Selected reaction monitoring of bacteria-related peptides was applied for the first time to identify bacteria including Staphylococcus aureus and Escherichia coli. This study presents an accurate means for bacterial identification in food matrixes using MS. The analysis time is less than 90 min and the minimum concentration of E. coli detected was 5 × 10 CFU/mL. The interaction between bacteria and the magnetic nanoparticles was electrostatic and nonspecific, in contrast to immunoassays which require specific antibodies. The targeted peptide analysis focuses on the bacterial markers, thus significantly simplifying the analysis and leading to an accurate identification of bacteria.

摘要

确保食品安全需要一种快速可靠的方法来检测食源性病原体。质谱已被证明是一种强大的工具,可以对纯细菌物种进行分类。然而,由于微生物信号的抑制,来自食物背景的基质干扰可能会导致错误的结果。因此,开发一种用于从食物样品中富集细菌和鉴定标记物的方法是很有用的。在特定条件下(pH 值为 4.5),使用磁性氧化锆纳米粒子从苹果汁/生菜中浓缩添加的微生物。使用纳升液相色谱-质谱法进行细菌鉴定。首次应用与细菌相关的肽的选择反应监测来鉴定包括金黄色葡萄球菌和大肠杆菌在内的细菌。本研究使用 MS 为食物基质中的细菌鉴定提供了一种准确的方法。分析时间小于 90 分钟,检测到的大肠杆菌最小浓度为 5×10 CFU/mL。与需要特定抗体的免疫测定相比,细菌与磁性纳米粒子之间的相互作用是静电的和非特异性的。靶向肽分析侧重于细菌标记物,因此大大简化了分析,实现了对细菌的准确鉴定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/064b/9296202/661f7fd5de2c/jfda-27-02-575f1.jpg

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